a Examples TIRF images of dispersed ND (left) and T2D α-cells (right) expressing Pppg-NPY-EGFP, after equilibration in the indicated glucose concentrations. Scale bar 2 µm. b Average exocytosis as function of ambient glucose concentration for dispersed ND (black) and T2D (red) α-cells as in a. For ND, n = 13 cells/3 donors at 1 mM, 8 cells/2 donors at 3 mM, 13 cells/2 donors at 7 mM, 15 cells/3 donors at 10 mM, and 10 cells/2 donors at 20 mM). For T2D, n = 7 cells/2 donors at 1 mM), 9 cells/2 donors at 3 mM, 10 cells/2 donors at 7 mM, 10 cells/2 donors at 10 mM, and 7 cells/2 donors at 20 mM. Data are presented as mean values ± SEM. c Docked granules (average granule density) as function of ambient glucose concentration for ND (black, n = 32 cells/5 donors at 1 mM, 32 cells/5 donors at 3 mM, 32 cells/5 donors at 7 mM, 32 cells/5 donors at 10 mM, and 32 cells/5 donors at 20 mM) and for T2D cells (red, n = 11 cells/2 donors, 11 cells/2 donors at 3 mM, 17 cells/2 donors at 7 mM, 16 cells/3 donors at 10 mM, and 11 cells/2 donors at 20 mM). Data are presented as mean values ± SEM. In b, c
p-values in black or red for comparisons as indicated by a line in ND group or T2D group respectively (two-tailed t-test). d Average rate of docking (granules becoming immobilized in the TIRF plane) as function of the ambient glucose concentration in the same cells as in b. Data are presented as mean values ± SEM. e Cumulative time course (upper), total exocytosis (middle), and initial density of docked granules (lower) during K+-stimulated exocytosis in dispersed ND α-cells bathed in 1 mM (30 cells/6 donors, yellow), 7 mM (38 cells/6 donors, blue) or 10 mM glucose (71 cells/14 donors, black). Stimulation was from 10 to 50 s. Data are presented as mean values ± SEM. f As in e, but for dispersed T2D α-cells. n = 27 cells/5 donors at 1 mM, 24 cells/3 donors in 7 mM, and 33 cells/6 donors in 10 mM glucose. In e, f, p-values in black for comparisons as indicated by a line, or in red comparing ND and T2D for the same condition (oneway ANOVA, Fisher posthoc test).